Collectively migrating tumor cells have been recently implicated in enhanced metastasis of epithelial malignancies.In oral squamous cell carcinoma(OSCC),av integrin is a crucial mediator of multicellular clustering an...Collectively migrating tumor cells have been recently implicated in enhanced metastasis of epithelial malignancies.In oral squamous cell carcinoma(OSCC),av integrin is a crucial mediator of multicellular clustering and collective movement in vitro;however,its contribution to metastatic spread remains to be addressed.According to the emerging therapeutic concept,dissociation of tumor clusters into single cells could significantly suppress metastasis-seeding ability of carcinomas.This study aimed to investigate the anti-OSCC potential of novel endostatin-derived polypeptide PEP06 as a clusterdissociating therapeutic agent in vitro.Firstly,we found marked enrichment ofαv integrin in collectivelyinvading multicellular clusters in human OSCCs.Our study revealed that metastatic progression of OSCC was associated with augmented immunostaining of av integrin in cancerous lesions.Following PEP06treatment,cell clustering on fibronectin,migration,multicellular aggregation,anchorage-independent survival and colony formation of OSCC were significantly inhibited.Moreover,PEP06 suppressed av integrin/FAK/Sre signaling in OSCC cells.PEP06-induced loss of active Src and E-cadherin from cell-cell contacts contributed to diminished collective migration of OSCC in vitro.Overall,these results suggest that PEP06 polypeptide 30 inhibiting av integrin/FAK/Src signaling and disrupting E-cadherin-based intercellular junctions possesses anti-metastatic potential in OSCC by acting as a cluster-dissociating therapeutic agent.展开更多
目的探讨绿茶提取物(green tea extract,GTE)对不同的人口腔鳞癌细胞株的抗肿瘤效应及其相关机制。方法体外培养人舌鳞癌CAL-27细胞株、人舌鳞癌SCC-25细胞株、人口腔上皮癌KB细胞株,应用MTT法检测GTE对细胞增殖的影响,筛选出敏感细胞株...目的探讨绿茶提取物(green tea extract,GTE)对不同的人口腔鳞癌细胞株的抗肿瘤效应及其相关机制。方法体外培养人舌鳞癌CAL-27细胞株、人舌鳞癌SCC-25细胞株、人口腔上皮癌KB细胞株,应用MTT法检测GTE对细胞增殖的影响,筛选出敏感细胞株,用流式细胞术检测GTE对CAL-27细胞周期的影响,采用蛋白芯片技术检测GTE对CAL-27细胞株蛋白表达的影响,并应用Western blot检测周期蛋白依赖性激酶4(cyclin-dependent kinases,CDK4)、周期蛋白依赖性激酶6(cyclin-dependent kinases,CDK6)、磷酸化3-磷酸肌醇依赖性蛋白激酶-1(phospho-3-phosphoinositide-dependent protein kinase-1,p-PDK1)蛋白表达情况。结果与对照组比较,50、100、200、400μg/m L GTE作用于CAL-27细胞抑制率升高,100、200、400μg/m L GTE作用于KB细胞抑制率升高,25、50、100、200、400μg/m L GTE作用于SCC-25细胞抑制率亦升高,差异均有统计学意义(P<0.01),并呈剂量依赖性。流式细胞术显示:与对照组比较,50μg/m L GTE介导CAL-27细胞G2/M期阻滞(P<0.05);100μg/m L GTE能够介导CAL-27细胞S期及G2/M期阻滞(P<0.01),G0/G1期细胞减少(P<0.01)。应用蛋白芯片技术共分析107种蛋白,应用GTE处理后,CAL-27细胞共有13种蛋白发生明显变化。应用Western blot技术确认了25、50、100μg/m L GTE作用于CAL-27细胞后p-PDK1、CDK4、CDK6蛋白表达,药物浓度越高,抑制率越高,差异有统计学意义(P<0.05)。结论 GTE能够抑制多种类型人口腔鳞癌细胞株的增殖,敏感细胞株为CAL-27。GTE主要影响表皮生长因子受体(EGFR)和Notch信号传导通路,并通过以上信号传导通路影响细胞周期相关蛋白表达水平的变化,导致细胞周期S期及G2/M期阻滞。展开更多
基金funded by the National Natural Science Foundation of China(grant Nos.81730012 and 81673426)the Grant of Republic Bashkortostan for Young Scientists(grant No.26 GR).
文摘Collectively migrating tumor cells have been recently implicated in enhanced metastasis of epithelial malignancies.In oral squamous cell carcinoma(OSCC),av integrin is a crucial mediator of multicellular clustering and collective movement in vitro;however,its contribution to metastatic spread remains to be addressed.According to the emerging therapeutic concept,dissociation of tumor clusters into single cells could significantly suppress metastasis-seeding ability of carcinomas.This study aimed to investigate the anti-OSCC potential of novel endostatin-derived polypeptide PEP06 as a clusterdissociating therapeutic agent in vitro.Firstly,we found marked enrichment ofαv integrin in collectivelyinvading multicellular clusters in human OSCCs.Our study revealed that metastatic progression of OSCC was associated with augmented immunostaining of av integrin in cancerous lesions.Following PEP06treatment,cell clustering on fibronectin,migration,multicellular aggregation,anchorage-independent survival and colony formation of OSCC were significantly inhibited.Moreover,PEP06 suppressed av integrin/FAK/Sre signaling in OSCC cells.PEP06-induced loss of active Src and E-cadherin from cell-cell contacts contributed to diminished collective migration of OSCC in vitro.Overall,these results suggest that PEP06 polypeptide 30 inhibiting av integrin/FAK/Src signaling and disrupting E-cadherin-based intercellular junctions possesses anti-metastatic potential in OSCC by acting as a cluster-dissociating therapeutic agent.
文摘目的探讨绿茶提取物(green tea extract,GTE)对不同的人口腔鳞癌细胞株的抗肿瘤效应及其相关机制。方法体外培养人舌鳞癌CAL-27细胞株、人舌鳞癌SCC-25细胞株、人口腔上皮癌KB细胞株,应用MTT法检测GTE对细胞增殖的影响,筛选出敏感细胞株,用流式细胞术检测GTE对CAL-27细胞周期的影响,采用蛋白芯片技术检测GTE对CAL-27细胞株蛋白表达的影响,并应用Western blot检测周期蛋白依赖性激酶4(cyclin-dependent kinases,CDK4)、周期蛋白依赖性激酶6(cyclin-dependent kinases,CDK6)、磷酸化3-磷酸肌醇依赖性蛋白激酶-1(phospho-3-phosphoinositide-dependent protein kinase-1,p-PDK1)蛋白表达情况。结果与对照组比较,50、100、200、400μg/m L GTE作用于CAL-27细胞抑制率升高,100、200、400μg/m L GTE作用于KB细胞抑制率升高,25、50、100、200、400μg/m L GTE作用于SCC-25细胞抑制率亦升高,差异均有统计学意义(P<0.01),并呈剂量依赖性。流式细胞术显示:与对照组比较,50μg/m L GTE介导CAL-27细胞G2/M期阻滞(P<0.05);100μg/m L GTE能够介导CAL-27细胞S期及G2/M期阻滞(P<0.01),G0/G1期细胞减少(P<0.01)。应用蛋白芯片技术共分析107种蛋白,应用GTE处理后,CAL-27细胞共有13种蛋白发生明显变化。应用Western blot技术确认了25、50、100μg/m L GTE作用于CAL-27细胞后p-PDK1、CDK4、CDK6蛋白表达,药物浓度越高,抑制率越高,差异有统计学意义(P<0.05)。结论 GTE能够抑制多种类型人口腔鳞癌细胞株的增殖,敏感细胞株为CAL-27。GTE主要影响表皮生长因子受体(EGFR)和Notch信号传导通路,并通过以上信号传导通路影响细胞周期相关蛋白表达水平的变化,导致细胞周期S期及G2/M期阻滞。
